Isolation and continuous cultivation of Trypanosoma theileri in media containing tissue culture fluids

Splitter, Earl J.; Soulsby, E. J. L.

doi:10.1016/0014-4894(67)90075-6

Cited by 17 publications

(4 citation statements)

References 10 publications

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“…For the blood flow stages of T. theileri (Trypomastigotes), RPMI medium 1640 with 10% FCS, supplemented with murine spleen cells as a feeder layer, was used (Verloo et al, 2000). In addition, it could be cultured in tissue culture fluid NCTC-109 (Splitter and Soulsby, 1967), 50% HMI-9 medium (Hirumi and Hirumi, 1989) supplemented with 20% FCS, 10% Serum, and 50% MDBK-conditioned media (Mott et al, 2011).…”

Section: Growth Media and Conditionmentioning

confidence: 99%

Molecular Identification of Trypanosoma theileri and Biology of Trypanosomes

Fentahun¹,

Paeshuyse²

2021

WVJ

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Trypanosoma theileri (T. theileri ) is a non-pathogenic, cosmopolitan, and commensal protozoa of cattle. The main objective of the current study was to investigate the biology and feasibility of T. theileri as a model candidate for the discovery of a novel drug. In the present study, the isolates of T. theileri obtained from the Institute of Tropical Medicine (ITM) in SDM 79 were cultivated at 26oC. Eight experiments with different inoculum and different times were grown. The growth curve was plotted to check the growth trends. The doubling time in the logarithmic phase was determined to be 17.43 hours. In addition, an experimental infection was done on a 3-month-old Holstein Friesian calf to isolate the blood-streaming shape; however, it was not successful after the blood buffy coat smear and PBMC culture in RPMI 1640 and HMI 9. Furthermore, the viability was determined by quantitative colorimetric Resazurin assay in 96-well fluorescence Microplates containing 0.4 to 2.4 mM of Resazurin. On the other hand, the response to Pentamidine (1-100 ng/mL) showed a strong negative correlation between the fluorescence signal and the highest Pentamidine concentration. IC50 was 9.25 ng/mL. Genomic DNA was extracted using the phenol-chloroform method. The gradient PCR amplification using T. theileri specific PCR (Tth625-PCR) primers was detected at 465 base pair (bp). In addition, the full-length 18S rDNA sequence was detected at 730 bp. In the silico analysis using common anti-trypanosome drug targets, no significant similarity could be found on either the DNA or the protein level. Nevertheless, homologous sequences have been identified among the drug targets for Ornithine decarboxylase. Therefore, the analysis might show the possibility of using T. theileri as a model for the search of new drugs once they have entire genome sequences. Analysis of the whole genome and transcriptome indicated a phylogenetic relationship between T. theileri and other pathogenic trypanosomes which can be the basis for novel drug development.

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Section: Growth Media and Conditionmentioning

confidence: 99%

Molecular Identification of Trypanosoma theileri and Biology of Trypanosomes

Fentahun¹,

Paeshuyse²

2021

WVJ

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“…The presence of blood cells, the temperature of cultivation and the type of culture vessel used appear to be important [38]. Cultivation at room temperature results in multiplication in the crithidial stage, and only in a few instances has the organism been cultivated at 37°C in the trypanosome stage [39]. Attempts by Herbert [38] to obtain continuous culture of T. theileri at 37°C were unsuccessful.…”

Section: Diagnosis and Confirmation Of Ttheilerimentioning

confidence: 99%

“…Later, T.theileri in the trypanosome form has been reported as a contaminant in cultures, at 37 o C, of normal bovine fetal kidney cells or blood lymphocytes and lymph node cells of cattle with lymphatic leukemia or viral diarrhea. Tissue culture fluids, supplemented with newborn calf serum and blood or blood products, are suitable for the isolation and continuous cultivation of T.theileri at 37°C [39]. Cultivation of the trypanosomes at 37 o C has been reported in non-defined or partially defined monophasic liquid media containing erythrocytes or erythrocyte products, in embryonal chicken eggs and in various types of cell cultures [40].…”

Section: Diagnosis and Confirmation Of Ttheilerimentioning

confidence: 99%

Molecular detection of Trypanosoma theileri and a new Trypanocidal drug, a review

Fentahun¹

2020

jlsb

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Introduction. Trypanosoma theileri (T.theileri) is apathogenic, cosmopolitan and commensal protozoa of cattle. Despite apathogenic in healthy, but not in stressed cattle; it's getting recent attention as a tool to tackle pathogenic ones. These days, researchers are giving due attention to study the biology and feasibility of T.theilerito use it as a model candidate for novel drug discovery. In addition, in-silico analysis using common antitrypanosome drug targets couldn't show significant similarity both at DNA and protein level. Nevertheless, homologous sequences have been identified among drug targets for Ornithine decarboxylase. This indicated the possibility to consider T.theileri as a model to search novel drugs once having whole genome sequences. The SDM 79 is an appropriate medium to cultivate at 26 o C, without CO 2 . Gradient PCR amplification has been used to detect using T. theileri. The specific primer (Tth625) which reveals 465 bp amplification product and also the full length 18S ribosomal DNA sequence of T.theileri DNA detectable at 730 base pairs are commonly used. Whole genome and transcriptome analysis can show the phylogenetic relationship between T.theileri and other pathogenic Trypanosomes which can be the basis for novel drugs development. Aim. The purpose of this paper is toreview the nature of Trypanosoma theileri, its molecular identification and also using its apathogenic nature as an opportunity to discover anew Trypanocidal drug.

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“…Normal kan muayenesinde, düşük parazitemi dolayısıyla, saptanamayan T. theileri'yi, sı-ğırlarda görebilmek için kültür metodlarının kullanılması önerilmektedir (5,8,22,27,32,35).…”

Section: Introductionunclassified

TÜRKİYE'DE SIĞIRLARDAN TRYPANOSOMA THEiLERi (LAVERAN, 1902)'NiN iZOLASYONU VE KÜLTiVASYONU

İncı¹,

Sayin²

1995

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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Isolation and cultivation of Trypanosoma theileri (Laveran, 1902) GirişTrypanosoma theileri (Kinetoplastorida, Trypanosomatidae) Laveran, 1902 kamçılı bir protozoon olup sığırların perifer kanında plazma içinde yaşar (9). Her kıtada bir çok ülkede görüldüğü için kozmopolit bir parazit olarak tanımlanır 04,19,23,34).Trypanosoma theileri oldukça büyüktür. Uzunluğu genellikle 60-70ji arasında değişir 04,19,23,24,31,34,36). Lanset şeklinde olan parazitin vücudunun arka ucu uzun ve sivri, kinetoplast arka uctan uzakta ve içeride yer alır, dalgalı zarı belirgin, flagellum uzun ve uç kısnu serbesttir 09,23,24,31,34).Sığırla-rın kanında bu parazitin trypomastigot ve epimastigot şekillerine, çeşitli doku ve lenf yumrularında, uzunluğuna ikiye bölünerek çoğalan, epimastigot şekillerine rastlanılmıştır (7,9,16,17,19,37,39).Başta Tabanus ve Haematopota olmak üzere Tabanidae ailesine bağlı çeşitli soylarda yer alan sokucu sineklerle bir sığırdan diğerinenakledilir01, 14,19,23,31,34).Trypanosoma theileri çok yaygın olmakla beraber genellikle patojen olmayan bir tür olarak tanımlanır

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Isolation and continuous cultivation of Trypanosoma theileri in media containing tissue culture fluids

Cited by 17 publications

References 10 publications

Molecular Identification of Trypanosoma theileri and Biology of Trypanosomes

Molecular Identification of Trypanosoma theileri and Biology of Trypanosomes

Molecular detection of Trypanosoma theileri and a new Trypanocidal drug, a review

TÜRKİYE'DE SIĞIRLARDAN TRYPANOSOMA THEiLERi (LAVERAN, 1902)'NiN iZOLASYONU VE KÜLTiVASYONU

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